Yin Xiongwei, Baldoni Irene, Adams Erwin, Van Schepdael Ann
Department of Pharmaceutical and Pharmacological Sciences, Pharmaceutical Analysis, KU Leuven, 3000 Leuven, Belgium.
Molecules. 2024 Dec 19;29(24):5993. doi: 10.3390/molecules29245993.
Analyzing and detecting endogenous amino acids in blood is of crucial importance for the diagnosis of medical conditions and scientific research. Considering the lack of UV chromophores in most of these analytes and the presence of several interfering substances in plasma, the quantification of quite a few amino acids and related compounds presents certain technical challenges. As a blank plasma matrix lacking these endogenous substances does not exist, the surrogate matrix method is used, as well as isotopic internal standards for calibration, to ensure the accuracy and reliability of the study. Method validation was conducted for 48 target analytes, giving the following results: linearity (R at least 0.99), limit of quantification (from 0.65 to 173.44 μM), precision (intra-day and inter-day RSD for LQC ranged from 3.2% to 14.2%, for MQC from 2.0% to 13.6%, and for HQC from 1.6% to 11.3%), accuracy, recovery, and stability of the method (all complied with the guidelines). To further investigate the applicability of this method to large-scale sample analysis, the method was successfully applied to the analysis of amino acids in plasma samples collected from 20 control individuals, demonstrating its wide application scope for clinical diagnosis and metabolic research.
分析和检测血液中的内源性氨基酸对于疾病诊断和科学研究至关重要。鉴于大多数这些分析物缺乏紫外发色团，且血浆中存在多种干扰物质，相当多氨基酸及相关化合物的定量分析存在一定技术挑战。由于不存在缺乏这些内源性物质的空白血浆基质，因此采用替代基质法以及同位素内标进行校准，以确保研究的准确性和可靠性。对48种目标分析物进行了方法验证，结果如下：线性（R至少为0.99）、定量限（0.65至173.44 μM）、精密度（LQC日内和日间RSD范围为3.2%至14.2%，MQC为2.0%至13.6%，HQC为1.6%至11.3%）、准确度、回收率和方法稳定性（均符合指导原则）。为进一步研究该方法对大规模样本分析的适用性，该方法成功应用于对20名对照个体采集的血浆样本中的氨基酸分析，证明了其在临床诊断和代谢研究中的广泛应用范围。
